• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.347-353
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• 1997

This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

• Parasites, Hosts and Diseases
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• v.35 no.1
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• pp.25-30
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• 1997

Gymnophclloides seoi is a human intestinal trematode prevalent on southwestern islands in Korea. In the present study, we investigated whether G. seoi metacercariae can grow and develop into adults by in vitro cultivation. The metacercariae were obtained from naturally infected oysters, and cultured in uitro for 5 days under three conditions; $37^{\circ}C/5%{\;}CO_2,{\;}41^{\circ}C/8%{\;}CO_2,{\;}or{\;}41^{\circ}C/15%{\;}CO_2$, in NCTC 109 complete media containing 20% FBS and 1% antibiotics-antimycotics. The degree of worm growth and development was compared with that grown in uiuo of C3H mice. The length of the worms cultivated in uitro was $200-300{\;}\mu\textrm{m}$, not significantly different from metacercariae, whereas the length of the worms recovered from C3H mice was significantly larger, $300-400{\;}\mu\textrm{m}$. The worms produced eggs when grown in C3H mice or cultured in vitro for 2 days under $41^{\circ}C/8%{\;}CO_2{\;}or{\;}41^{\circ}C/5%{\;}CO_2$, but not when cultured under 37$^{\circ}C/5%{\;}CO_2$. Among the in vitro conditions, $41^{\circ}C/15%{\;}CO_2$ was best for egg Production, although the number of eggs was about half of worms obtained from C3H mice. In conclusion, in vitro cultivation of G. semi metacercariae into egg-pioducing adults was partially successful under culture conditions of $41^{\circ}C/5%{\;}CO_2{\;}or{\;}41^{\circ}C/8%{\;}CO_2$.

• The Korean Journal of Mycology
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• v.51 no.2
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• pp.69-80
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• 2023

The cultivation conditions of shiitake mushrooms (Lentinula edodes) influence the production and quality of fruiting bodies. We conducted this study to improve the productivity and quality of shiitake mushrooms by modifying the cultivation conditions. Two types of spawns (sawdust and liquid spawn) were used, and corn flour was used as a nutritional source for the sawdust medium. A blue light-emitting diode (LED; 300 lux) was also used instead of a white LED during the incubation period. Sanbaekhyang was used as the experimental variety. When using corn flour, the mycelial growth rate increased 1.1 to 2.7 times the growth rate of the control up to 21 days of incubation, and the weight loss rate of the media was also higher. Mushroom productivity increased 1.2 times when the liquid spawn was used compared to when the sawdust spawn was used, and the blue LED also increased fruiting body production by 1.1 times compared to the white LED. Mushroom productivity increased when the liquid spawn was used, and the blue LED also increased fruiting body production. Fruiting body weight and the size of the cap were greater when sawdust spawn was used. The fruiting body weight and the stipe diameter were greater when the blue LED was used. Taste analysis showed that the saltiness increased when corn flour was used, and the sourness increased when the blue LED was used.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.179-187
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• 2011

This review describes the characteristics of various unculturable soil bacteria, successfully-cultivating examples of those bacteria, and the diverse factors to be considered for successful cultivation. Most importantly, the selection of proper media is very important because unculturable bacteria demand different types of nutrients at various concentrations of substrates, nitrogens and phosphorus. To develop a new medium to successfully culture unculturable bacteria from soil, molecular ecological studies should be combined together. The inoculum size on a plate is also important: less than 50 bacterial cells are recommended to be plated on a single culture plate. The environmental factors such as pH and salt concentration of the medium need to be adjusted as similar as possible to mimic the original soil environments, and the trial of the various temperatures and extended period of cultivation are better. Since one cannot simply tell about which one was unculturable among a great number of colonies grown on a newly developed medium, some suitable detection methods and fast identification methods are required. Many soil bacteria live with cooperation one another in their communities, so that enrichment such as coculture of using other bacterial metabolites and subsequent pure cultures can also guarantee successful cultivation of the previously uncultured bacteria in soil. Here, this review will discuss for the future perspectives to culture the unculturable soil bacteria.

• Economic and Environmental Geology
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• v.34 no.4
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• pp.345-354
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• 2001

The effects on arsenic geochemistry of indigenous microorganisms isolated from an area contaminated with high concentration of arsenic were investigated. Arsenite exerted higher inhibitory effects on the microbes' growth than arsenate. During incubation of the microbes in an arsenate-spiked medium over 24 hours, decrease in microbial growth was observed as arsenate content increased. Arsenate of 150 mM or over apparently inhibited cell growth. However, further incubation for up to 4 days in the high arsenate concentration medium resulted in cell growth, implying that the microorganisms adjusted their biochemical functions to detoxify arsenic and maintain growth. Two types of microbes were observed during 20 hours to reduce arsenate to arsenite in solution through a detoxification mechanism. As well, decrease in the total arsenic content occurred over a 4-day incubation with the same microbes in an arsenate-spiked medium. Therefore it is suggested that microorganisms can influence arsenic speciation in natural settings and this may be applied to efficient bioremediation of arsenic-contaminated sites.

• Microbiology and Biotechnology Letters
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• v.10 no.2
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• pp.87-93
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• 1982

The cultural conditions of Mucor plumbeus FRI 0007 were investigated for the maximum production of felt and lipid. It was found that the lower the pH and the higher the incubation temperature, the higher accumulation of the felt and lipid. Shake culture rendered higher lipid accumulation and lower felt accumulation than static culture. Maximum production of felt and lipid content were 47.8 g/$\ell$ and 50.73%, respectively, when the organisms were static-cultured at a temperature of 37$^{\circ}C$ and pH of 3.5 for 25 days latroscan thinchrographic analysis showed that the higher amount of triglyceride was obtained when static-cultured at a low pH. Fatty acid composition of the microbial lipid was affected by the incubation temperature, types of nitrogen source and speed of agitation: lower degree of saturation was observed as the incubation temperature decreased and the speed of agitation increased. Fatty acids of monoglyceride and diglyceride were mainly palmitic and oleic acids and those of triglycerides were mainly palmitic, oleic acids.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.4
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• pp.493-498
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• 2006

The optimal culture conditions of Monascus purpureus MMK2 for production of red pigment were investigated in submerged culture. Monascus purpureus MMK2 showed a maximal production of red pigment in the medium containing of 3.0% wheat flour, $0.15%\;NaNO_3,\;0.25%\;Na_2HPO_4\;12H_2O$ and $0.15%\;MgSO_4\;7H_2O$. The optimal culture conditions of temperature and initial pH were $30^{\circ}C$ and 6.5, respectively. The red pigment production reached to a maximal level at 7th day of cultivation.

• Journal of Korean Society of Forest Science
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• v.97 no.3
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• pp.285-290
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• 2008

This study was carried out to obtain an optimal liquid culture condition for Sarcodon aspratus mycelia. Among various basal culture media the GYS (glucoe-yeast extract-soytone) medium was the best for mycelial growth. The appropriate temperature was $25^{\circ}C$. Starch, maltose or glucose was excellent carbon sources for the mycelial culture, compared to others tested. As nitrogen nutrients, soytone and $NH_4-N$ were the best organic and inorganic nitrogen sources, respectively. Moreover, the optimal concentration of soytone was 3 g per one-liter medium. In addition, we also found that alanine, $(NH_4)H_2PO_4$, and nicotinic acid were the best aminoacid, phosphorus salt, and vitamin, respectively. When all optimal conditions described above were applied to culture medium, we were able to produce 5.7 g dry weight of S. aspratus mycelia per one-liter liquid medium within 20 days.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.84-86
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• 2014

Mass production of biocontrol agent is an essential step for its commercial use. Media composition and culture conditions for production of Bacillus amyloliquefaciens SKU-78, a potential biocontrol agent against bacterial wilts, were optimized by a flask culture. Low cost media combining nitrogen and carbon sources were tested. Maximum cell growth (> $2{\times}10^9$ CFU/ml) was obtained in a medium of 5% soy flour combined with 3% corn starch after 24 h cultivation. The optimum initial pH, temperature and shaking speed was 5.5, $30^{\circ}C$ and 150-250 rpm, respectively. Fermentation of SKU-78 was scaled up in 30 L fermenter and the profiles of cell density, pH, dissolved oxygen and spore formation were recorded. After 8 h lag phase, exponential growth occurred and reached at maximum viable cell number ($1.2{\times}10^{11}$ CFU/ml) after 20 h. The SKU-78 strain grown in a low cost medium exhibited the high suppression of bacterial wilts. The results indicate that SKU-78 strain can be produced in a low cost medium and provide a basis for scaling up to industrial level.

• The Microorganisms and Industry
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• v.15 no.2
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• pp.33-39
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• 1989

본고에서는 주로 방선균의 선택분리 과정을 시료(substrate)선정, 시료 전처리(pretreatment), 배지, 배양조건, 콜로니 선정 등으로 나누어 살펴보고자 한다.